Department of Virology and Immunology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland.
J Biosci. 2009 Dec;34(6):927-40. doi: 10.1007/s12038-009-0107-2.
We studied the expression of inducible heat shock protein (HSP27, HSP72) and multidrug-resistance protein (MRP) in co-cultures of human colon carcinoma cell spheroids obtained from different grades of tumour with normal human colon epithelium, myofibroblast and endothelial cell monolayers. We also measured the influence of recombinant human transforming growth factor beta1 (rhTGF-beta1) and camptothecin (CPT-11), added as single agents or in combination, on the levels of the HSPs, MRP, interleukin (IL)-6 and nitric oxide (NO). An immunoblotting analysis with densitometry showed that rhTGF-beta1 and/or CPT-11 increased HSP27, HSP72 and MRP expression in tumour cells and myofibroblasts, as well as in co-cultures compared with appropriate controls. By contrast, in colonic epithelium, inhibition of HSPs and MRP was comparable with that of the control. In endothelial cells, HSP72 was undetectable. Direct interaction of colon tumour spheroids with normal myofibroblasts caused a significant, tumour-grade dependent increase in IL-6 production. Production of IL-6 was significantly lowered by rhTGF-beta1 and/or CPT-11. Tumour cell spheroids cultivated alone produced larger amounts of NO than normal cells. In co-culture, the level of the radical decreased compared with the sum of NO produced by the monocultures of the two types of cells. rhTGF-beta1 and/or CPT-11 decreased NO production both in tumour and normal cell monocultures and their co-cultures. In conclusion, direct interactions between tumour and normal cells influence the expression of HSP27, HSP72 and MRP, and alter IL-6 and NO production. rhTGF-beta1 and/or CPT-11 may potentate resistance to chemotherapy by increasing HSP and MRP expression but, on the other hand, they may limit tumour cell spread by decreasing the level of some soluble mediators of inflammation (IL-6 and NO).
我们研究了诱导热休克蛋白（HSP27、HSP72）和多药耐药蛋白（MRP）在不同分级的人结肠癌肿瘤细胞球与正常结肠上皮、肌纤维母细胞和内皮细胞单层共培养物中的表达。我们还测量了重组人转化生长因子β1（rhTGF-β1）和喜树碱（CPT-11）作为单一药物或联合应用对 HSPs、MRP、白细胞介素（IL）-6 和一氧化氮（NO）水平的影响。免疫印迹分析和密度测定显示，rhTGF-β1 和/或 CPT-11 增加了肿瘤细胞和肌纤维母细胞以及共培养物中 HSP27、HSP72 和 MRP 的表达，与适当的对照相比。相比之下，在结肠上皮细胞中，HSPs 和 MRP 的抑制作用与对照相似。在内皮细胞中，HSP72 无法检测到。结肠癌肿瘤球与正常肌纤维母细胞的直接相互作用导致 IL-6 产生显著的、依赖肿瘤分级的增加。rhTGF-β1 和/或 CPT-11 显著降低了 IL-6 的产生。单独培养的肿瘤细胞球产生的 NO 量大于正常细胞。在共培养中，与两种类型细胞的单核培养物产生的 NO 总和相比，自由基的水平降低。rhTGF-β1 和/或 CPT-11 降低了肿瘤和正常细胞单核培养物及其共培养物中 NO 的产生。总之，肿瘤细胞与正常细胞的直接相互作用影响 HSP27、HSP72 和 MRP 的表达，并改变 IL-6 和 NO 的产生。rhTGF-β1 和/或 CPT-11 可能通过增加 HSP 和 MRP 的表达增强化疗耐药性，但另一方面，它们可能通过降低某些炎症可溶性介质（IL-6 和 NO）的水平来限制肿瘤细胞的扩散。
